Development of Col.1-TAZ transgenic mice.

<p>(A) Linearized structure of the Col.1–TAZ construct used for transgenic expression. The coding sequence of mouse TAZ was placed under the pro-α1 (I) collagen promoter. The 350 bp EcoRI-BamHI fragment was used as a probe in Southern blot analysis. The two-way arrows indicate the primers used for PCR genotyping. Both DNA probe and the 243 bp PCR product encompass the junction between the Flag tag and the 5′end of the TAZ reading frame. (B) Southern blot of the genomic DNA of a litter showing a 1.5 kb band product corresponding to the transgene. (C) Total protein extracts isolated from calvariae of either Col.1–TAZ mice or the WT littermate were separated by SDS-PAGE and blotted (Western blot) using an anti-Flag or anti-TAZ antibody. A band of 50 kDa, corresponding to the transgene, was detected by an anti-Flag antibody in the extracts from Col.1–TAZ mice but not from the WT littermate. Expression of the transgene was further confirmed by increased expression levels of TAZ in Col.1–TAZ mice detected by the TAZ antibody compared to the WT littermate.